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Light Sheet Box

The Light Sheet Box enables you to build an advanced fluorescence microscopy system that illuminates only a thin slice of your specimen. This technique allows for fast, gentle, and high-contrast 3D imaging of biological samples.

What is Light Sheet Microscopy?​

Light sheet microscopy illuminates the sample perpendicular to the detection axis using a thin sheet of laser light. Only the focal plane is illuminated while all other regions remain dark, which:

  • Reduces photobleaching and phototoxicity
  • Increases imaging speed dramatically
  • Provides excellent optical sectioning
  • Enables 3D reconstruction through z-stack imaging

How it Works​

A laser beam is expanded and shaped using a cylindrical lens to create a thin light sheet that passes through the sample perpendicular to the microscope's optical axis. The objective lens views the illuminated plane from the side, capturing only fluorescence from the thin illuminated section. By moving the sample through the light sheet in small increments, you can acquire a complete 3D volume.

What You'll Learn​

  • Principles of light sheet illumination
  • Fluorescence microscopy fundamentals
  • 3D image acquisition and reconstruction
  • Cylindrical lens optics and beam shaping
  • Motorized stage control for 3D scanning
  • Sample preparation for light sheet imaging

Experiments in this Section​

  • Light Sheet Fluorescence Microscope - Build a complete light sheet microscopy system
  • Light Sheet Sample Preparation - Preparing and mounting specimens
  • Light Sheet Calibration - Alignment and optimization procedures

Key Capabilities​

  • Fast 3D imaging of transparent specimens
  • Gentle illumination minimizes photobleaching
  • High-contrast optical sectioning
  • Automated XYZ scanning
  • Simultaneous bright-field alignment
  • Image stack reconstruction

Required Components​

  • Laser diode (488nm for fluorescence excitation)
  • Biconvex lens (f'=25mm)
  • Cylindrical lens (f'=100mm) for sheet generation
  • Mirror for beam steering
  • LED satellite matrix for bright-field alignment
  • Electronic Z-stage for camera focus
  • Electronic XYZ-stage for sample movement
  • Mainboard with USB connection
  • PS4 controller for stage control
  • Infinity objective (10x)
  • Sample objective (4x with RMS mount)
  • Probe chamber for sample mounting
  • Emission filter
  • Camera with tube lens
  • UC2 cubes and baseplates

Ideal for developmental biology, neuroscience research, and anyone interested in cutting-edge 3D microscopy techniques!